CRISPR/Cas9 gene editing to generate Drosophila LexA lines in secondary school classes

Anne E. Rankin,Elizabeth Fox,Townley Chisholm,Nicole Lantz,Arjun Rajan,William Phillips,Elizabeth Griffin,Jaekeb Harper, Christopher Suhr,Max Tan,Jason Wang,Alana Yang,Ella S. Kim, Naa Kwama A. Ankrah, Praachi Chakraborty, Alistair C. K. Lam, Madeleine E. Laws,Jackson Lee, Kyle Park,Emily Wesel, Peter H. Covert,Lutz Kockel,Sangbin Park,Seung K. Kim

biorxiv(2023)

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摘要
Genome editing in vivo with CRISPR/Cas9 generates powerful tools to study gene regulation and function. We developed CRISPR-based methods that permitted secondary school student scientists to convert Drosophila GAL4 lines to LexA lines. Our novel curricula implement a new donor strain optimizing Homology-assisted CRISPR knock-in (HACK) that simplifies screening using light microscopy. Successful curricula adoption by a consortium of schools led to the generation and characterization of 16 novel LexA lines. This includes extensive comparative tissue expression analysis between the parental Gal4 and derived LexA lines. From this collaboration, we established a workflow to systematically generate LexA lines from frequently-used GAL4 lines. Modular courses developed from this effort can be tailored to specific secondary school scheduling needs, and serve as a template for science educators to innovate courses and instructional goals. Our unique collaborations highlight that resources and expertise harnessed by university-based research laboratories can transform experiential science instruction in secondary schools while addressing research needs for the community of science.
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